NOTCH Signaling Is Activated through Mechanical Strain in Human Bone Marrow-Derived Mesenchymal Stromal Cells

Figure 6

Knockdown of NOTCH1 and NOTCH2 genes in hMSC-TERT-AP-1 cells. (a) Immunoblotting of whole cell lysates of hMSC-TERT-AP-1 cells with antibodies against NOTCH1 and NOTCH2. Full-length NOTCH (NFL), transmembrane and intracellular part of NOTCH (NTM), and housekeeping protein vinculin. (b) hMSC-TERT-AP-1 cells were stably transduced with lentiviral vectors expressing shRNAs against NOTCH1 (shN1_1 and shN1_2). Relative mRNA expression of NOTCH1, HES1, HEY1, HES6, FOS, and PTGS2 is shown. Results are given as mean of two independent and normalized to an empty vector (dashed line). (c) Immunoblotting of whole cell lysates with antibodies against NOTCH2 after transfer of shN2 and nontargeting shRNA (ctr). (d) Relative mRNA expression of NOTCH2, HES1, HEY1, HES6, FOS, and PTGS2 after transfer of shN2 compared to nontargeting shRNA (dashed line). Two-tailed unpaired -test was used for statistical analysis (, , and ). qPCR data were obtained from technical triplicates.

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